Neonatal Fc receptor FcRn is involved in intracellular transport of the Fc fusion protein aflibercept and its transition through retinal endothelial cells

AbstractRetinal endothelial cells (REC) likely contribute to the clearance of intravitreally injected IgG. Because this is of high relevance to the pharmacokinetic assessment of the widely used therapeutic Fc fusion protein aflibercept, we studied its transport through immortalized bovine REC (iBREC) in detail. For shuttling of IgG or Fc fusion proteins like aflibercept, endothelial cells use the highly conserved neonatal Fc receptor (FcRn) also expressed in iBREC where it is down regulated by serum depletion. Therefore, we focused on studying intracellular localization and transport of aflibercept under conditions affecting its interaction with the FcRn. Intracellular localization of aflibercept was assessed by Western-blot analyses of subcellular protein fractions or by immunofluorescence staining. After uptake in a temperature-dependent process, aflibercept co-localized with early endosomes, which harbor FcRn. Similar amounts of aflibercept were co-extracted with proteins from membranes/organelles irrespectively of the amount of FBS in the culture medium. Lowering the concentration of FBS resulted in a strong, but reversible association with cytoskeletal proteins suggesting a block in intracellular transport. In accordance with this finding, aflibercept's transport through an iBREC monolayer grown on porous membrane inserts was markedly delayed in the absence of FBS in the culture medium indicating that aflibercept is taken up but not exocytosed under these conditions. Transcytosis of aflibercept was also strongly delayed by inhibition of phosphatidylinositol 3-kinase with LY294002, which affects FcRn-mediated IgG transport. A similar inhibition of aflibercept's transport was observed with IgG-binding proteins (i.e. protein A or protein G) that block interaction between FcRn and aflibercept. Interfering with aflibercept's binding to the FcRn with protein A (or protein G) or the inhibitory FcRn-specific monoclonal antibody 1G3 resulted in a reduced amount of intracellular aflibercept. Taken together, our results strongly suggest that FcRn is involved in transport of aflibercept through REC in vitro

Susceptibility of Atlantic cod Gadus morhua, halibut Hippoglossus hippoglossus and wrasse (Labridae) to Aeromonas salmonicida subsp. salmonicida and the possibility of transmission of furunculosis from farmed salmon Salmo salar to marine fish

The possibility of transmission of Aeromonas salmonicida subsp. salmonicida from Atlantic salmon Salmo salar to cod Gadus morhua, halibut Hippoglossus hippoglossus, and wrasse (Labridae) was studied in both laboratory cohabitation and injection challenge trials, and in a field situation during an outbreak of furunculosis in Atlantic salmon in 2 sea farms containing both cod and salmon. A, salmonicida subsp, salmonicida was isolated from cod, halibut and wrasse, but only occasionally. The injection challenge studies with A, salmonicida subsp. salmonicida were carried out on cod and halibut. Despite challenge with 4.6 x 10(3), 4.6 x 10(4) and 4.6 x 10(5) CFU (colony-forming units) of the bacterium, only 1 halibut died, whereas no mortality occurred in the cod groups. In cohabitation challenge trials in which cod, halibut and wrasse were kept together with infected salmon, 1 cod and 1 halibut died, with subsequent isolation of A, salmonicida subsp. salmonicida from their kidneys. In a group of salmon challenged in the same way with infected salmon cohabitants, all the salmon died. In the field study, there were high mortalities of salmon due to furunculosis. However, A, salmonicida subsp. salmonicida was isolated from only 1 of the wrasse used as 'cleaner fish' for the salmon. A. salmonicida was not isolated from cod during the field study. The results suggest that A, salmonicida subsp. salmonicida can be transmitted from salmon and cause disease in cod, halibut and wrasse, but that this is a relatively rare event

Effects of dietary menthol-rich bioactive lipid compounds on zootechnical traits, blood variables and gastrointestinal function in growing sheep

Background The present study aimed at investigating the influence of 90% menthol-containing plant bioactive lipid compounds (PBLC, essential oils) on growth performance, blood haematological and biochemical profile, and nutrient absorption in sheep. Twenty-four growing Suffolk sheep were allotted into three dietary treatments: Control (without PBLC), lower dose of PBLC (PBLC-L; 80 mg/d) and higher dose of PBLC (PBLC-H; 160 mg/d). Sheep in all groups were fed meadow hay ad libitum plus 600 g/d of concentrate pellets for 28 d. Results Average daily gain was not affected by treatment. Feeding of PBLC increased hay and total feed intake per kg body weight (P < 0.05). Counts of total leucocytes, lymphocytes and monocytes were not different among treatments. However, neutrophil count decreased (P < 0.05) in PBLC-H with a similar trend in PBLC-L (P < 0.10). Concentrations of glucose, bilirubin, triglycerides, cholesterol, urea and magnesium in serum were not different among sheep fed different doses of PBLC. However, serum calcium concentration tended to increase in PBLC-H (P < 0.10) and serum concentrations of aspartate & asparagine (P < 0.01) and glutamate & glutamine (P < 0.05) increased linearly with increasing PBLC dose. In ruminal epithelia isolated from the rumen after killing, baseline conductance (Gt; P < 0.05) and short-circuit current (Isc; P < 0.01) increased in both PBLC groups. Ruminal uptakes of glucose and methionine in the presence of Na+ were not affected by the dietary PBLC supplementation. In the absence of Na+, however, glucose and methionine uptakes increased (P < 0.05) in PBLC-H. In the jejunum, Isc tended to increase in PBLC-H (P < 0.10), but baseline Gt was not affected. Intestinal uptakes of glucose and methionine were not influenced by PBLC in the presence or absence of Na+. Conclusion The results suggest that menthol-rich PBLC increase feed intake, and passive ion and nutrient transport, the latter specifically in the rumen. They also increased serum concentrations of urea precursor amino acids and tended to increase serum calcium concentrations. Future studies will have to show whether some of these findings might be commonly linked to a stimulation of transient receptor potential (TRP) channels in the gastrointestinal tract

Morphology of Starch Particles along the Passage through the Gastrointestinal Tract in Laboratory Mice Fed Extruded and Pelleted Diets

Simple Summary Starch is the main carbohydrate source in most lab mouse diets. Its properties are influenced by feed processing. This determines how easily accessible it is to enzymatic digestion in the gastrointestinal tract of animals. In previous studies we have shown that there are differences between pelleted and extruded forms of a maintenance diet fed to mice regarding digestibility and microbiome. To complement these findings, the present study presents a morphological study of the starch particles throughout the passage along the gastrointestinal tract of C57BL/6J mice fed either pellets or extrudate. Samples were stained with Lugol's iodine and examined via stereomicroscope and scanning electron microscope. Starch granules in the pelleted diet are mostly intact and compact, thus autoenzymatic digestion in the small intestine is less efficient than in the more accessible starch granules from the extruded diet. For both diet forms, starch accumulation in the caecum was observed, suggesting selective retention of praecaecally undigested starch for microbial fermentation. These findings allow for unique insights in murine starch digestion that are important to understand the digestive physiology of this species. Diet processing impacts on starch properties, such as the degree of starch gelatinization. This affects digestibility, as shown in laboratory mice fed either a pelleted or an extruded diet. In the present study, the morphology of starch particles throughout the digestive tract of mice was visualized. Thirty-two female C57BL/6J mice were used for a feeding trial. They were fed a commercial maintenance diet for laboratory mice, which was available in pelleted and extruded form, for seven weeks. The mice were sacrificed after the feeding period, and chyme samples were collected from five sites (stomach, anterior and posterior small intestine, caecum, colon). Samples of diets, chyme and faeces were analyzed via stereomicroscopy (stained with Lugol's iodine) and scanning electron microscopy (SEM). The starch granules appeared more compact in the pelleted diet, showing first signs of degradation only in the small intestine. The caecum content of both diets group was intensively stained, particles as well as fluid phase, indicating that it contained mainly starch. The SEM pictures of caecum content showed abundant bacteria near starch particles. This suggests selective retention of prae-caecally undigested starch in the murine caecum, likely the site of microbial fermentation

Seroprevalence of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum Infections in German Horses

There are limited data on Lyme borreliosis (LB), a tick-borne disease caused by the Borrelia burgdorferi sensu lato complex, in horses. Seropositivity is not necessarily associated with clinical disease. Data on seropositivity against Borrelia burgdorferi and Anaplasma phagocytophilum in German horses are sparse. Therefore, serum samples from horses (n = 123) suspected of having Lyme borreliosis and clinically healthy horses (n = 113) from the same stables were tested for specific antibodies against Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. The samples were screened for antibodies against Borrelia burgdorferi (ELISA and an IgG line immunoblot assay). Furthermore, the samples were examined for antibodies against B. burgdorferi and Anaplasma phagocytophilum with a validated rapid in-house test (SNAP® 4Dx Plus® ELISA). The clinical signs of suspect horses included lameness (n = 36), poor performance (n = 19), and apathy (n = 12). Twenty-three percent (n = 26) of suspect horses and 17% (n = 18) of clinically healthy horses were seropositive for having a Borrelia burgdorferi sensu lato infection (p = 0.371), showing that the detection of specific antibodies against B. burgdorferi alone is not sufficient for a diagnosis of equine LB. Anaplasma phagocytophilum seropositivity and seropositivity against both pathogens was 20%/6% in suspect horses and 16%/2% in the clinically healthy population, showing only minor differences (p = 0.108). Unspecific testing for antibodies against B. burgdorferi without clinical suspicion of Lyme borreliosis is not recommended since the clinical relevance of seropositivity against Borrelia burgdorferi sensu lato remains to be elucidated

Phylogenetic relationships in the southern African genus Drosanthemum (Ruschioideae, Aizoaceae)

Background. Drosanthemum, the only genus of the tribe Drosanthemeae, is widespread over the Greater Cape Floristic Region in southern Africa. With 114 recognized species, Drosanthemum together with the highly succulent and species-rich tribe Ruschieae constitute the 'core ruschioids' in Aizoaceae. Within Drosanthemum, nine subgenera have been described based on flower and fruit morphology. Their phylogenetic relationships, however, have not yet been investigated, hampering understanding of monophyletic entities and patterns of geographic distribution. Methods. Using chloroplast and nuclear DNA sequence data, we performed network- and tree-based phylogenetic analyses of 73 species of Drosanthemum with multiple accessions for widespread species. A well-curated, geo-referenced occurrence data set comprising the 134 genetically analysed and 863 further accessions was used to describe the distributional ranges of intrageneric lineages and the genus as a whole. Results. Phylogenetic inference supports nine clades within Drosanthemum, seven of which group in two major clades, while the remaining two show ambiguous affinities. The nine clades are generally congruent to previously described subgenera within Drosanthemum, with exceptions such as (pseudo-) cryptic species. In-depth analyses of sequence patterns in each gene region were used to reveal phylogenetic affinities inside the retrieved clades in more detail. We observe a complex distribution pattern including widespread, species-rich clades expanding into arid habitats of the interior (subgenera Drosanthemum p.p., Vespertina, Xamera) that are genetically and morphologically diverse. In contrast, less species-rich, genetically less divergent, and morphologically unique lineages are restricted to the central Cape region and more mesic conditions (Decidua, Necopina, Ossicula, Quastea, Quadrata, Speciosa). Our results suggest that the main lineages arose from an initial rapid radiation, with subsequent diversification in some clades.Raw data, code, analysis output, and species occurrence The zip file contains a ReadMe file and 4 folders: 1_main_data_and_results (the files used to produce the figures in the main text); 2_ML_phylogenetics (raw data, code, and analysis output of ML phylogenetic analyses); 3_MJ_networks (raw data [SNP/sequence motive recoded DNA alignment matrices], and output of median-joining network analyses)

Модификация поверхности имплантов органическими функциональными группами

Работа направлена на разработку модификации поверхности аддитивно полученных скэффолдов на основе титанового сплава Ti6Al4V с аренедиазониевыми тозилатами и металл-органическими каркасами для улучшения биосовместимости и процесса остеоинтеграции. Целью исследования является разработка методов ковалентной модификации поверхностей скэффолдов с использованием арендиазоний тозилатов и металлоорганических каркасов для повышения выживаемости имплантатов в организме человека.The work is aimed at developing a surface modification of additively obtained scaffolds based on a Ti6Al4V titanium alloy with arenediazonium tosylates and metal-organic frameworks to improve biocompatibility and osseointegration. Aim of research to develop methods for covalent modification of scaffold surfaces using arenediazonium tosylates and metal-organic frameworks for better implant survival in the human body

Value transmission in primary schools: are teachers’ acculturation orientations a moderator?

The transmission of human values to primary school pupils is key, which is acknowledged in curricula in a variety of cultural contexts worldwide. The present study presents data from the multicultural and multilingual region of Bolzano in Northern Italy (n = 422 pupils in k = 30 classrooms). In addition to class teachers’ values as predictors of their pupils’ values we investigated whether teachers’ acculturation orientations would strengthen the relationship between teachers’ and children’s values, thereby acting as moderators. We focused on the opposing acculturation orientations of integration-transformation versus exclusion. A multilevel analysis showed that teachers’ conservation values (tradition, conformity, and security) significantly predicted their pupils’ conservation values, and that teachers’ openness to change values (self-direction, stimulation, and hedonism) significantly predicted their pupils’ openness to change values. This indicates successful value transmission in the classroom. Teachers’ self-transcendence (benevolence and universalism) and self-enhancement (achievement and power) values did not significantly predict pupils’ values. As expected, teachers’ acculturation orientations were related to their values, but they did not play a role in predicting pupils’ values. Implications for value transmission in the school context and for acculturation research are discussed

Interlacing properties and bounds for zeros of 2ϕ1 hypergeometric and little q-Jacobi polynomials

Please read abstract in the article.http://link.springer.com/journal/111392017-05-30hb201

Comparative Analysis of Intestinal Helminth Infections in Colic and Non-Colic Control Equine Patients

All around the world, intestinal helminths constitute one of the most prevalent life-long occurring infections and re-infections affecting all horse age groups. A range of parasite species among strongyles, ascarids, and tapeworms is known to have the potential to cause colic in horses. However, there is a lack of current scientific evidence on the actual relevance of helminth infection levels in the context of colic in horses kept during prevailing epidemiological conditions. Thus, a prospective case-control study on the occurrence of intestinal helminths in a total of 620 mainly adult equine clinic patients was conducted to investigate the association between colic and helminth infection. For each horse, a range of copromicroscopic, serological, and clinical data was obtained, in addition to a questionnaire on relevant anamnestic data, including previous anthelmintic treatment and husbandry. Using a FLOTAC-based copromicroscopic diagnosis, the highest infection rates were seen for strongyles (41.8%), followed by Anoplocephala perfoliata and Parascaris spp. (both 0.8%), with no significant difference between the two study groups. Employing a real-time PCR a 1.1% S. vulgaris DNA prevalence was found. Considerably higher seroprevalences were observed using S. vulgaris and A. perfoliata ELISAs, with 32.3% and 10.7%, respectively. It was noteworthy that no association concerning either serologic status was encountered with colic status. The shedding of strongyle eggs was associated with a 1.8-times increased risk of S. vulgaris seropositivity. Recent anthelmintic treatment was associated with the onset of colic, as animals who had received an anthelmintic during the previous week had a 2.4-times higher risk of signs of colic compared to those who had been treated at least eight weeks prior. Another noteworthy observation was that ponies were significantly less often affected by colic than warmbloods. The high S. vulgaris and considerable A. perfoliata seroprevalences encountered in this investigation should prompt veterinarians, farm managers, and horse owners to maintain consequent and effective worm control measures